Antipyretic agent

ABSTRACT

3-O-(A-L-MYCAROSYL)-8-HYDROXYERYTHRONOLIDE B IS USEA PHARMACOLOGICALLY ACTIVE SUBSTANCE HAVING BLOOD FUL AS AN ANTIPYRETIC AGENT. THE COMPOUND IS PREPARED BY THE FERMENATION OF 8-HYDROXYERYTHRONOLIDE B WITH STREPTOMYCES ERYTHREUS NRRL 3887 IN SUITABLE NUTRIENT MEDIA.

United States Patent 3,740,425 ANTIPYRETIC AGENT Jerry Roy Martin, Waukegan, Ill., assignor to Abbott Laboratories, North Chicago, Ill.

No Drawing. Original application Dec. 18, 1970, Ser. No. 99,705, now Patent No. 3,684,794. Divided and this application Mar. 9, 1972, Ser. No. 233,340

Int. Cl. A61k 21/00 US. Cl. 424-181 1 Claim ABSTRACT OF THE DISCLOSURE 3-O-(a-L-mycarosyl)-8-hydroxyerythronolide B is useful as an antipyretic agent. The compound is prepared by the fermentation of 8-hydroxyerythronolide B with Streptomyces erythreus NRRL 3887 in suitable nutrient media.

REFERENCE TO RELATED APPLICATIONS This application is a divisional application of copending application bearing Ser. No. 99,705, filed Dec. 18, 1970, now U.S. Pat. 3,684,794.

DISCLOSURE OF THE INVENTION This invention relates to the novel compound 3-O-(a-L- mycarosyl)-8-hydroxyerythronolide B which has useful pharmodynamic activity. More specifically, the compound of this invention has an antipyretic effect as demonstrated in standard animal pharmacological procedures. For example, an antipyretic effect is observed in yeast fevered rats at oral dosages of 100-200 mg. kg.

The compound of this invention has the following structural formula:

The compound of this invention is prepared by the fermentation of a nutrient media comprising a carbohydrate energy source such as a monosaccharide, e.g., glycose and corn starch; a source of nitrogen such as soy flour, corn steep liquor, ammonium nitrate and the like; and S-hydroxyerythronolide B, which media has been seeded with a culture of Streptomyces erythreus NRRL 3887. It is also preferable that the nutrient media comprise a buffering agent to moderate the pH. Such agents are dihydrogen, phosphate and alkaline earth carbonates, e.g., calcium carbonate.

The S-hydroxyerythronolide B is obtained according to the method described in the United States patent application entitled Erythronolide B Derivatives, filed concurrently with this application on behalf of Paul Kurath, which filing date and serial number will here be inserted as soon as they are made known.

The compound of this invention may be administered orally or parenterally in conventional dosage forms such as tablets, capsules, injectables and the like, which incorporate 3 O-(a-L-mycarosyl)-8-hydroxyerythronolide B 3,740,425 Patented June 19, 1973 were prepared in a medium consisting of (in grams per liter) glycose monohydrate (Cerelose), 15.0; soy bean meal, 15.0; and calcium carbonate, CaCO 1.0. The cultures were incubated at 32 C. for 72 hours on a rotary shaker. The seed was added at a level of 35% (v./v.) into 500 ml. Erlenmyer flasks containing 50 ml. of a fermentation medium consisting of the following components (in grams per liter): corn starch, 15.0; soya fluff flour, 20.0; corn steep, 5.0 CaCO 1.0; and soy bean oil (Edsoy) 50.0. The medium was adjusted to pH 6.6 with sodium carbonate and autoclaved at 120 for 30 minutes at 15 pounds pressure before inoculation. The fermentation flasks were incubated at 32 on a rotary shaker (260 r.p.m.) for 48 hours. One gram of finely divided 8-hydroxyerythronolide B was equally distributed among 40 fermentation flasks and incubation with shaking was continued for 120 hours.

The fermentation beer was clarified by the addition with stirring of an equal volume of an aqueous solution of 10% Zinc sulfate followed by an equal volume of 0.5 N sodium hydroxide. A filter aid, Dicalite, was added and the mixture was stirred for 5 minutes. The mixture was filtered and the clear filtrate of pH 6.6 was collected. The filtrate was extracted twice with equal volumes of ethyl acetate. The combined ethyl acetate extract was washed twice with water and dried over anhydrous sodium sulfate. Concentration in vacuo gave 1.23 grams of viscous yellow-brown oil which solidified on standing. The solid material was dissolved in a small amount of chloroform and added to the top of a column (3.0 x 35 cm.) of silica gel (Brinkman, -325 mesh) prepared in chloroform. Elution with increasing concentrations of methanol in chloroform gave fractions containing only a material with R, 0.29 by thin layer chromatography (Silica Gel G, ethanol-chloroform, 1:10 v./v.). These fractions were collected and the solvent removed in vacuo to give 797 mg. of a light yellow oil. The oil was dissolved in methanol and treated with an equal weight charcoal (DarcoG-60). Crystallization from methanol-water gave 327.5 mg. of 3-a-L-mycarosyl-8-hydroxyerythronolide B as fine white needles, MP. 205- 208 C.

When the compound of this invention was administered orally at a dosage of 200 mg./kg. of body weight to mice who had a yeast induced fever, the results were that at one hour following administration, the fever was reduced by 69.6% and three hours after administration, the reduction was 28.2%; at mg./kg., the reduction was 15.9 and 5.5% respectively.

I claim:

1. A method of reducing fever in warm-blooded animals comprising administering to said warm-blooded animal of from 100-200 mg./kg. of body weight of the compound 3-O-(a-L-mycarosyl)-8-hydroxyerythronolide B.

References Cited UNITED STATES PATENTS 3,629,232 12/1971 Jones 260-2105.

JEROME D. GOLDBERG, Primary Examiner 

